巢式聚合酶链反应扩增外周血吞噬细胞内外伤寒沙门菌Vi基因

目的：明确外周血吞噬细胞内外伤寒沙门菌Ｖｉ基因检测在伤寒诊断中的意义。方法：高速离心沉淀，低渗溶解红细胞法分离８５例临床拟诊伤寒患者外周血吞噬细胞（内）及吞噬细胞外（血清中）伤寒沙门菌，巢式聚合酶链反应扩增伤寒沙门菌Ｖｉ基因；并与单管套式聚合酶链反应扩增粒细胞内伤寒沙门菌Ｈ基因、巢式聚合酶链反应扩增单核细胞内伤寒沙门菌Ｖｉ基因、１次聚合酶链反应扩增血清中伤寒沙门菌Ｈ基因比较。结果：巢式聚合酶链反应

DETECTION OF SALMONELLA TYPHI VI GENE IN PERIPHERAL BLOOD SERUM AND MACROPHAGE BY NESTED PCR AMPLIFICATION

Objective:To clarify the significance of detecting Salmonella typhi Vi gene in peripheral blood serum and macrophages in diagnosis of typhoid fever.Methods:Detection of S.typhi Vi gene in peripheral blood and macrophages from 85 patients with clinical typhoid fever by nested PCR amplification method,S.typhi H gene in granulocytes by single tube nested PCR,S.typhi Vi gene in mononuleocytes by nested PCR and S.typhi H gene in serum by PCR.Results:There were no significant differences (P>0.05)in the positivity rate among S.typhi Vi gene in peripheral blood serum and macrophages(64.7%),S.typhi Vi gene in mononucleocytes(60.0%)and S.typhi H gene in granulocytes(57.6%);but the positivity rate of them was significantly higher(P<0.01)than that of S.typhi H gene in serum(32.9%).Conclusions:The nested PCR amplification technique is a simple,time saving,sensitive and specific method for detecting S.typhi Vi gene in peripheral blood serum and macrophages.It may be used as an ideal parameters for laboratory diagnosis of typhoid fever.